OCCURRENCE 

OF 

MICROSCOPIC  CRYSTALS 

IN  THE 

VERTEBRAS 

OF  THE 

TOAD. 


H,  CARRINGTON  BOLTON. 


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Ufl* 


[Reprinted  from  the  Proceedings  of  the  American  Association  for  the  Advance¬ 
ment  of  Science,  Yol.  XXVIII,  Saratoga  Meeting,  August,  1879.] 


On  the  Occurrence  of  Microscopic  Crystals  in  the  Verte¬ 
bra  of  the  Toad  ( Bufo  Americanus).  By  H.  Carrington 
Bolton,  of  Hartford,  Conn.  With  a  Note  by  Alexis  A. 
Julien,  of  New  York. 

The  microscopic  crystals  contained  in  the  capsules  attached  to 
the  spinal  nerve  of  the  frog  have  been  examined  by  many  Histol¬ 
ogists.  They  were  first  noticed,  we  believe,  by  the  learned  Flem¬ 
ish  physician,  Blasius  1  in  the  latter  part  of  the  seventeenth 
century.  Swann,  in  his  Comparative  Anatomy  of  the  Nervous 
System ,2  gives  figures  of  them  without  any  detailed  description, 
and  Wagner3  refers  to  the  crystals  as  follows:  “The  ganglia 
admit  of  being  very  readily  demonstrated,  lying  upon  the  sides  of 
the  vertebral  column  ;  they  are  situated  near  those  small  white 
vesicles  which  protrude  by  becoming  swollen,  chiefly  during  the 


1  Anat.  Animalium,  Amst.  p.  291, 1681. 

2  Plate  VII,  figs.  5  and  6. 

8  Comp.  Anat.  Vertebrates,  part  I.,  p.  151. 


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p 37415 


2 


MICROSCOPIC  CRYSTALS  ; 


spring  of  the  year,  and  contain  numerous  microscopic  calcareous 
crystals.”  They  were  also  observed  by  Stannius  and  by  Prof. 
Owen,  who,  however,  added  little  to  our  knowledge  of  them. 

In  1852  Dr.  Jeffries  Wyman  prepared  a  memoir  on  the 
“Anatomy  of  the  Nervous  System  of  Rana  Pipiens,”  which  is 
published  in  the  fifth  volume  of  the  Smithsonian  Contributions 
to  Knowledge,  (1853).  In  this  paper  he  states  that  the  “  ciystal 
capsules  are  subject  to  some  variety,  but  exist  on  the  trunks  of 
all  the  true  spinal  nerves,  the  hypoglossus  included,  and  invest 
them  more  or  less  completely  at  their  exit  from  the  spinal  canal, 
occupying  the  space  between  the  transverse  processes  of  two 
adjoining  vertebrae.  The  capsule  is  a  very  thin  membrane,  some¬ 
times  having  a  diameter  of  2  to  3  lines  ;  its  surface  is  minutely 
lobulated  in  appearance,  which  results  from  the  existence  of 
numerous  partitions  within,  forming  many  small  cavities,  which 
communicate,  though  not  very  freely,  with  each  other.  The 
contents  of  these  pouches  are  vast  numbers  of  exceedingly  minute 
crystals  of  a  somewhat  oval  form,  pointed  at  their  extremities, 
and  comparable  in  shape  to  a  lemon  seed,  but  sometimes  pre¬ 
senting  well  defined  angles.” 

Dr.  Wyman  failed  to  confirm  Wagner’s  statement  that  the 
capsules  are  more  swollen  in  the  spring  of  the  year,  having,  he 
says,  “detected  them  of  nearly  equal  dimensions  at  all  seasons,” 
and  he  also  differs  from  Stannius  in  regarding  them  as  character¬ 
istic  of  the  adult  animal. 

Dr.  W}7 man  then  remarks:  “Dissections  of  other  than  Rani- 
form  Batrachians  have  not  led  to  their  detection  ;  I  have  not 
found  them  in  Siren ,  Menobranckus ,  Bufo  or  Salamandra ,  but 
have  seen  them  in  Rana  fontinalis ,  Rana,  halecina  and  Rana 
palustris.  They  seem  to  be  in  these  species  constant  appendages 
to  the  spinal  nerves.” 

Notwithstanding  this  positive  statement  of  so  distinguished 
and  accurate  an  observer,  the  writer  has  found  the  microscopic 
crystals  in  the  toad  in  great  abundance,  and  differing  little  in  form 
and  general  appearance  from  those  occurring  in  the  frog.  Speci¬ 
mens  of  the  common  toad  ( Bufo  Americanus) ,  collected  both  in 
the  autumn  and  in  the  spring  were  dissected,  and  in  eveiw  instance 
the  contents  of  the  spinal  canal  examined  under  a  \  inch  object¬ 
ive,  disclosed  the  white  matter  as  a  mass  of  minute  crystals. 

On  treating  the  crystals  on  a  glass  slide  with  acetic  acid  they 


BY  H.  CARRINGTON  BOLTON. 


3 


dissolve  with  effervescence,  and  the  addition  of  oxalate  of  ammo¬ 
nium  to  this  solution  produces  a  milkiness  visible  to  the  unaided 
eye,  which  under  the  microscope  appears  to  be  formed  of  minute 
ovoid  masses  characteristic  of  precipitated  calcium  oxalate. 

When  the  calcareous  crystals  are  placed  under  the  microscope 
in  a  drop  of  water,  they  exhibit  in  a  marked  manner  the  phenom¬ 
ena  known  as  Brownian  motions  ;  the  larger  crystals  remain  at 
rest  but  the  smaller  ones  having  a  length  of  about  milli¬ 

metre  maintain  a  constant  restless  motion.  That  similar  crys¬ 
tals  from  the  spinal  canal  of  the  frog  exhibit  this  phenomenon  is 
well  known  to  histologists.4 

The  resemblance  of  the  crystals  from  the  toad  to  those  from 
the  frog  is  very  close,  but  a  careful  examination  shows  that  the 
former  average  somewhat  smaller  than  the  latter.  Slides  contain¬ 
ing  crystals  from  Rana  and  Bufo,  mounted  in  Canada  balsam, 
were  submitted  for  accurate  measurements  to  my  friend,  Mr. 
Alexis  A.  Julien,  to  whom  I  am  greatly  indebted  for  a  pains¬ 
taking  examination. 

Mr.  Julien  reports  as  follows  : 

“  The  crystals  appear  perfectly  colorless  with  sharp  outlines 
and  with  a  refractive  index  so  nearly  that  of  the  Canada  balsam 
in  which  they  are  mounted  that  it  is  difficult  to  distinguish  any 
but  their  outer  edges.  It  will  be  seen  b}r  the  measurements  given 
below  that  the  average  size  and  the  variation  in  size  of  the  Bufo 
cr}’stals  are  the  greater  of  the  two.  In  the  accompanying  sketch, 
Fig.  2,  d,  represents  one  of  the  larger  of  these  crystals,  exhib¬ 
iting  most  distinctly  the  highly  modified  edges.  In  both  the  Rana 
and  the  Bufo  crystals  many  were  observed  which  at  first  glance 
displayed  a  curved  or  spindle-shaped  outline  tapering  rather  sud¬ 
denly  toward  either  end  ;  some  of  these  are  shown  in  Figs.  1  and 
3.  They  are  often  of  considerable  size  and  by  careful  focusing 
these  curved  outlines  were  found  to  consist  of  a  succession  of 
straight  edges,  indicating  simply  a  more  highly  modified  form  of 
the  common  crystals.  On  the  whole  the  crystals  of  both  Rana 
and  Bufo  appear  to  me  to  differ  from  the  otoliths  of  aragonite 
enclosed  in  the  human  ear,  familiar  to  histologists,  and  to  re¬ 
semble  rather  highly  modified  rhombohedra  of  calcite  belonging, 
perhaps,  to  the  type  represented  in  Fig.  553  of  Dana’s  System  of 
Mineralogy.  No  twins  were  anywhere  observed.” 


4  Frey’s  Microscopical  Technology,  translated  by  Dr.  Geo.  R.  Cutter,  p.  101  (1872). 


4 


MICROSCOPIC  CRYSTALS  ;  BY  H.  CARRINGTON  BOLTON. 


Size  of  Crystals  infractions  of  millimetre. 


Bufo. 

Largest 
Smallest 
Common  size 


Length. 

0-033 

0-0005 

0-0034  to  0*025 


Breadth. 

0-0140 
0  0003 

0-001  to  0-01. 


liana. 

Largest  0*0229  0*0074 

Smallest  0-0007  0*0004 

Common  size  0-0068  0-0014 

[Otoliths  measure  about  0*012  mm.  long  by  0-008  mm.  broad.5] 

The  accompanying  figures  from  drawings  by  Mr.  Julien  show 
the  crystals  as  seen  with  a  magnifying  power  of  900  to  1008 
diameters,  under  the  camera  lucida. 

Fig.  1  shows  a  portion  of  the  field  seen  in  the  slide  of  Ran  a  crys¬ 
tals  and  Fig.  3  shows  the  same  in  that  of  Bufo,  with  the  usual 
intermixture  of  perfect  crystals  and  irregular  fragments.  Fig.  2 
contains  several  individual  crystals  enlarged  in  the  drawing. 


POSTSCRIPT.  — May,  1880. 


Since  preparing  the  foregoing  notes  Mr.  Simon  H.  Gage,  Instructor 
in  Physiology  at  Cornell  University,  has  had  the  kindness  at  my  sug¬ 
gestion  to  extend  the  examination  for  calcareous  crystals  to  other  Am¬ 
phibia.  He  reports  as  follows  : 

“Alcoholic  specimens  of  Siren  and  Menopoma  were  very  carefully  ex¬ 
amined,  but  no  crystals  could  be  found  either  in  the  neural  canal  or  on 
the  spinal  nerves.  A  fresh  Menobranchus  was  examined  with  the  same 
result.” 

Mr.  Gage  confirms  our  observation  as  to  the  occurrence  of  the  crystals 
in  Bufo  and  in  this  connection  remarks : 

“In  Rana  the  crystals  ou  the  nerves  are  beyond,  periplierad  ox,  the 
ganglion  and  seem  to  have  no  connection  with  those  lining  the  neural 
canal ;  while  in  Bufo  the  ganglion  is  completely  surrounded  by  a  cushion 
of  crystals  which  seem  to  be  merely  a  lateral  extension  through  the  in- 
tevertebral  foramina  of  the  general  cushion  lining  the  neural  canal. 

Dr.  Wyman  in  the  paper  quoted  makes  no  reference  to  the  crystals  in 
the  neural  canal  of  Rana.” 

Mr.  Gage’s  communication  is  accompanied  by  a  cross-section  of  the 
spinal  column  of  Bufo  showing  the  exact  position  of  the  cushion  of  crys¬ 
tals,  and  we  regret  that  circumstances  prevent  a  reproduction  of  his 
sketch. 

5  Dr.  J.  B.  St.  John  Roosa  in  Diseases  of  the  Ear,  p.  475  (1874). 


[Printed  at  the  Salem  Press,  June,  1SS0.] 


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